Transformation-mediated chromosome synthesis and replacement in eukaryotic cells
Authors: Alessandro L. V. Coradini; Cara B. Hull; Joshua Roemer; Daniel T. Lusk; Zachary Krieger; Ian M. Ehrenreich
Affiliation: University of Southern California, USC, Los Angeles, CA
Keywords: Comparative genomics & genome evolution
Genetic manipulation is one of the central strategies that biologists use to investigate the molecular underpinnings of life and its diversity. In recent years, the synthesis of chromosomes, known as synthetic genomics, has emerged as a new form of genetic manipulation. Megabase-sized chromosomes can now be generated from components synthesized de novo, obtained from naturally occurring genomes and other existing molecules, or a mixture of the two. Although de novo chromosome synthesis starting from oligonucleotides has been used in most synthetic genomics projects to date, this approach is too laborious and expensive to apply to many biological problems. To facilitate more widespread use of synthetic genomics, we developed an efficient method of chromosome synthesis and replacement in yeast based on the capture and reassembly of natural DNA segments. Here, I will describe the method and its application to questions about chromosome architecture and the genetic basis of phenotypic differences between strains, species, and potentially even genera.